转基因甘蔗BtG-2的T-DNA侧翼序列分析及其转化事件特异性检测

转基因甘蔗BtG-2是利用农杆菌介导法把Cry1Ac-2A-gna融合抗虫基因导入‘新台糖22号’的转基因甘蔗株系,具有良好的抗虫特性和农艺性状。为了明确转基因甘蔗BtG-2的分子特征及其检测方法,推进其生物安全性评价工作,以BtG-2的T2代为研究材料,利用Southern杂交检测外源基因在转基因甘蔗基因组内的拷贝数;利用染色体步移技术分离外源基因在甘蔗基因组中插入位点的侧翼序列,并建立了该转化体高效灵敏的特异性PCR检测方法。结果表明：Southern杂交检测证明外源T-DNA以单拷贝方式插入BtG-2株系;经过3次的热不对称巢式PCR扩增,获得外源基因T-DNA左边侧翼序列984 bp和右边侧翼序列705 bp;以这2个序列和相应的T-DNA的左右端序列分别设计3对检测引物对,建立了BtG-2株系的转化事件特异性PCR检测方法,扩增效率最高的引物对LS011/LA451和RS160/RA588分别扩增到440 bp和428 bp的特异片段。其中T-DNA左侧设计的LS011/LA451检测引物对扩增的灵敏度高、特异性强,能够在甘蔗BtG-2基因组DNA相对含量为0.1%的模板中检测出转基因目的成分,相当于9个单倍体基因组拷贝数。本研究完成了转基因株系BtG-2的分子特征及其转化事件特异性检测,为该转基因甘蔗及其衍生产品的检测和身份识别提供技术依据。     

香蕉新品种‘南天红’

香蕉新品种‘南天红’由‘南天黄’的自然突变选育而成。假茎平均高度2.59 m，幼树假茎外表面暗红色，抽蕾后转绿。雄花苞片外表面常为深红色，花苞卷曲，常不脱落；从种植到收获11 ~ 13个月，单株产量25 kg，品质好。中高抗香蕉枯萎病4号小种，采用配套的栽培技术可在所有香蕉种植区推广。     

The effects of various plant protection methods on the development of Zymoseptoria tritici and Cephalosporium gramineum,grain yield and protein profile

Septoria leaf blotch progresses rapidly, leading to the development of Zymoseptoria titici forms resistant to fungicides. Cephalosporium stripe is caused by Cephalosporium gramineum. The aim of this study was to evaluate the effectiveness of selected pesticides in limiting the symptoms of both diseases on winter wheat leaves, and to determine their influence on grain yield and the content and composition of protein fractions in wheat kernels. Propiconazoles were most effective in inhibiting the development of Septoria leaf blotch (symptoms were reduced from 54.7% to 78.6%). Strobilurins were less effective due to the presence of isolates with the G143A mutation. Symptoms of Cephalosporium stripe were rarely observed, and protective treatments did not reduce their severity. The highest content of grain protein (14.81%) was found in plants most intensely protected with the fungicides containing fenpropimorph, pyraclostrobin and epoxiconazole. The principal component analysis revealed that the plant protection method influenced the grain protein profile. The accumulation of HMW glutenins and α/β gliadins was mutually interrelated and higher in high-input treatments; control grain was characterized by close relationships between ω-gliadins, LMW glutenins, albumins and globulins, whereas low-input treatments influenced mostly γ-gliadins.     

S1核酸酶突变检测法的优化

优化了S1核酸酶突变检测体系,结果表明(1)扩增法比杂交法产生异源双链DNA更节省时间;(2)PCR产物可以直接作为突变检测的底物;(3)适当降低反应温度、适当增加反应缓冲液中的NaCl浓度、适当缩短反应时间可提高突变检测效果。     

梨芽变新品种—尖把王梨的选育

尖把王梨为尖把梨大果芽变,平均单果质量189g,最大单果质量280g,是一般尖把梨的2倍以上。部分果面有1～2条纵沟,果面黄绿色;果皮薄,果肉白色,肉质细腻多汁,石细胞较少;总糖含量为11.88%,总酸含量为0.92%,维生素C含量为5.81mg/100g,风味酸甜适口,香气浓,果心小,可食率高,品质上,是鲜食、冻制和加工兼用新品种。枝条、叶片也明显大于尖把梨,抗寒能力与尖把梨相同,是栽培梨中抗寒力最强的品种之一,适于辽宁、吉林、内蒙古等较寒冷地区发展。     

Preliminary results of using in vitro axillary and adventitious buds in mutation breeding of Chinese gooseberry

Summary Aseptically cultured shoots of Chinese gooseberry exhibited growth disorder and morphological aberrances, and some died after being exposed to sufficient gamma-ray irradiation. The death rate was dose dependant and the LD₅₀ was 80–90 Gy and 50–60 Gy respectively for cv. Hayward and clone 4. All petiole explants irradiated with gamma-ray could form calli as the control, but the rate of differentiation of adventitious shoots of the petiole explants decreased and was dependant on dose. Sensitivity of the shoot or petiole explants to gamma-ray irradiation varied with species. Gamma-ray irradiation did not deter either the 2-node segments from producing axillary shoots M₁, M₂, and M₃ or the advantitious shoots originating in the petiole explants and the M₃ shoots from forming advantitious roots. Therefore, using aseptically cultured axillary or adventitious buds for mutation breeding of Chinese gooseberry is feasible. A bacterium surviving in the explants lessened the efficiency of these two in vitro techniques in mutation breeding of Chinese gooseberry.Abbreviations IAA 3-indole acetic acid - IBA -indole butyric acid - MS Murashige & Skoog (1962)     

梨新品种‘花盖王’

 ‘花盖王’是从‘花盖梨’中选出的梨四倍体芽变新品种。该品种除保持‘花盖梨’的优质、丰产、抗寒等优良性状外，最突出特点是果实大，单果质量是‘花盖梨’的2倍多，最大为250 g。     

极早熟葡萄新品种‘90- 1’

 ‘90- 1’是从‘乍娜’的芽变中选出的葡萄新品种。该品种除保持母本的优质、丰产、适合保护地栽培等优良性状外, 其最突出的特点是极早熟, 果实6 月下旬成熟, 果实发育期仅35 d。     

狗牙根辐射诱变后代外部性状变异分析

对1997-2002年经过辐射诱变得到的狗牙根诱变后代的6个坪用性状指标的统计分析结果表明,1)不同的辐射诱变剂量处理的后代间坪用性状有显著差异,尤其在节间直径和密度上,6 800 rads处理的后代节间都比9 000 rads处理的后代粗,而密度是前者低于后者,2种剂量处理都可以显著降低草层高度;2)不同辐射诱变代数的后代间存在着差异,原始种源进行多代的辐射诱变,可以使得叶片显著变窄,草层高度显著降低,节间变细变长; 3)同一材料在相同辐射剂量诱变处理后匍匐茎不同节之间也存在显著的差异.     

秋水仙素处理日本矮紫薇种子变异的初步研究

用0.05％、0.10％、0.15％3种不同浓度秋水仙素和24、48、72 h 3个不同处理时间,对日本矮紫薇种子发芽性状和幼苗生长性状进行研究.结果表明:0.05％、0.10％、0.15％浓度秋水仙素处理日本紫薇种子24、48、72 h,随着秋水仙素浓度的增加和处理时间的延长其种子发芽率、发芽势、出苗率均有降低的现象,0.15％浓度秋水仙素溶液处理的日本矮紫薇种子72 h时出苗率低于10％.随着秋水仙素处理浓度和时间的延长,幼苗生长表现出矮化、根部变短、叶片变厚的趋势.该研究获得了3株矮化的变异植株及2株株型变异的幼苗,对日本矮紫薇突变体产生、筛选及良种选育提供了基础.